| Interleukin-1b ( IL-1b) is a potent stimulator of bone resorption whose gene is mapped to 2q14, and has been implicated in the pathogenesis of high bone turnover and osteoporosis. IL-1b, a prominent microglia-derived cytokine, caused oligodendrocyte death in coculture with astrocytes and microglia, but not in pure culture of oligodendrocytes alone. It also can cause nuclear export of a specific NCOR corepressor complex, resulting in derepression of a specific subset of nuclear factor-kappa-B (NFKB)-regulated genes2. Furthermore, Microenvironmental IL-1b and, to a lesser extent, IL-1a are required for in vivo angiogenesis and invasiveness of different tumor cells. Additional, the cooperation of IL-1b and PDGFB induces contractile-to-synthetic phenotype modulation of human aortic smooth muscle cells in culture. Moreover, the association with disease may be explained by the biologic properties of IL-1b, which is an important proinflammatory cytokine and a powerful inhibitor of gastric acid secretion. The Interleukin-1 beta, Human, BioAssay(TM) ELISA Kit is a sandwich ELISA for quantitative detection of human IL-1 beta in cell culture supernates, cell lysates, serum and plasma (heparin, EDTA). Detection Range: 3.91-250pg/ml Sensitivity: <0.15pg/ml Specificity: Recognizes native and recombinant human IL-1b. Test Principle: This Interleukin-1 beta, Human, BioAssay(TM) ELISA Kit is a solid-phase immunoassay specially designed to measure human IL-1 beta with a 96-well strip plate that is pre-coated with antibody specific for IL-1 beta. The detection antibody is a biotinylated antibody specific for IL-1 beta. The capture antibody is monoclonal antibody from mouse and the detection antibody is polyclonal antibody from goat. The kit contains recombinant human IL-1 beta with immunogen: Expression system for standard: E. coli, Immunogen sequence: A117-S269. To measure human IL-1 beta, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS buffer and add TMB. TMB is an HRP substrate and will be catalyzed to produce a blue color product, which changes into yellow after adding the acidic stop solution. The absorbance of the yellow product at 450nm is linearly proportional to Human IL1B in the sample. Read the absorbance of the yellow product in each well using a plate reader, and benchmark the sample wells readings against the standard curve to determine the concentration of human IL-1 beta in the sample. Kit Components: 143885A: Microtiter Strip, 1x96-well 143885B: Human IL-1 beta Standard, 2x1vial 143885C: Anti-human IL-1 beta (Biotin), 1x100ul (1:100 dilution) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x100ul (1:100 dilution) 143697: Sample diluent buffer, 1x30ml 143698: Antibody diluent buffer, 1x12ml 143699: ABC diluent buffer, 1x12ml 143700: TMB color developing agent, 1x10ml 143701: TMB stop solution, 2N H2SO4, 1x10ml 143702: Wash Buffer, 25X, 1x20ml Storage and Stability: Store *143885B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add 100ul of samples and standards and incubate the plate at 37C for 90 minutes or at RT for 2 hours. Do not wash. 2. Add 100ul biotinylated antibody and incubate the plate at 37C for 60 minutes or at RT for 90 minutes. Wash plate 3 times with Working Wash Buffer. 3. Add 100ul of ABC working solution and incubate the plate at 37C for 30 minutes or at RT for 40 minutes. Wash plate 5 times with Working Wash Buffer. 4. Add 90ul of TMB color developing agent and incubate the plate at 37C in dark for 15-25 minutes or at RT for 30 minutes. 5. Add 100ul TMB Stop Solution and read. |
